Green Fluorescent Protein Antibody

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Green Fluorescent Protein Antibody gallery image 1

Published customer image:
Sheep anti Green Fluorescent Protein antibody used for the identification of GFP tagged VEGF-A expressing cells by immunofluorescence.
Image caption:
HSV-1 infection drives transcriptional upregulation of VEGF-A.
(A) Representative image of a cornea from pVEGFA-GFP reporter mice at day 3 PI with HSV-1 strain McKrae stained for HSV-1 antigen (red), GFP transcriptional reporter for VEGF-A (green) and DAPI (blue) (B) GFP transcriptional reporter expression (green) with HSV-1 antigen (red) at 12 hours PI with HSV-1 strain McKrae. (C) Real time PCR of THCE cells at 12 hours PI with 3 pfu per cell HSV-1 strain McKrae with VEGF-A fold induction determined via the geometric means of VEGF-A induction relative to the housekeeping genes β-actin, TBP and PPIA (** p<0.01). (D) Real time PCR of VEGF-A transcription in 293 cells infected with 3 pfu per cell HSV-1 strain McKrae also determined via geometric mean relative to β-actin, TBP and PPIA (** p<0.01) and (E) Levels of VEGF-A by cytokine bead array in cytoplasmic extracts of 293 cells at the indicated times PI with 3 pfu per cell HSV-1 strain McKrae, expressed as pg of VEGF-A per μg of cytoplasmic protein extract. (F) Transcriptional activity of the proximal human VEGF-A promoter was measured using a luciferase vector with luciferase driven by the VEGF-A promoter (spanning base pairs -2018 to +50 bp relative to the transcription start site). Transfected 293 cells were infected with 3 pfu per cell of either wild-type HSV-1 strains; McKrae, KOS, or SC16 or with 3 pfu per cell HSV-2 and assayed for luciferase activity at the indicated time PI. Luciferase activity was normalized to the activity of uninfected 293 cells transfected with a promoterless luciferase vector, pGL3 (** p<0.01). All figures are representative figures from individual experiments with an n?=?3/group for A–D. Extracts from 3 culture wells per time point were pooled for E. Bars denote mean VEGF-A pg per μg of cytoplasmic protein ± SEM.

From: Wuest T, Zheng M, Efstathiou S, Halford WP, Carr DJJ (2011)
The Herpes Simplex Virus-1 Transactivator Infected Cell Protein-4 Drives VEGF-A Dependent Neovascularization.
PLoS Pathog 7(10): e1002278.

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Green Fluorescent Protein Antibody gallery image 2

Published customer image:
Sheep anti Green Fluorescent Protein antibody used for the identification of GFP tagged VEGF-A expressing cells by immunofluorescence.
Image caption:
The HSV-1 transactivator ICP4 is required for VEGF-A expression.
Human 293 cells were plated and infected with 3 pfu per cell HSV-1 strain McKrae. Fold induction of VEGF-A transcript expression was determined via geometric means of fold induction of VEGF-A transcript relative to the housekeeping genes β-actin, TBP and PPIA. (A) Inhibition of protein synthesis with cycloheximide (100 μg/mL) blocked upregulation of VEGF-A following HSV-1 infection while (B) inhibition of viral DNA synthesis with acyclovir (200 μM) had no significant effect. (C) ICP4 dependence was tested using human 293 cells infected with 3 pfu per cell of wild type HSV-1 KOS or KOS-derived mutants lacking ICP4 (ICP4- null virus) or the HSV-1 origin binding protein (OBP- null virus). VEGF-A transcript was measured via real time RT-PCR relative to β-actin, TBP and PPIA. (*p<0.05 relative to uninfected control cells). (D) Human 293 cells transfected with pVA8855 or the promoterless luciferase control plasmid, pGL3. Luciferase activity was measured at 12 hours PI with HSV-1 KOS, ICP4- null, or OBP- null virus. (E) VEGF-A mRNA levels in human 293 cells infected with 3 pfu per cell HSV-1KOS or an HSV-1 ICP0- null virus that fails to make the IE co-transactivator protein, ICP0; both viruses significantly up-regulated VEGF-A mRNA levels. (F) Mice lacking type I interferon responses due to deficiency in the type I interferon receptor (CD118-/-) were infected with HSV-1 KOS or HSV-1 ICP0- null virus. Corneas were harvested at 24 hours PI with 105 pfu of HSV-1 and analyzed for VEGF-A levels by cytokine bead array, expressed as pg of VEGF-A per mg of cornea wet mass. VEGF-A was induced by inoculation with HSV-1 KOS or HSV-1 ICP0- null virus (** p<0.01 relative to uninfected, scarified controls). (G) Reporter mice expressing GFP under the human VEGF-A promoter were analyzed for HSV-1 antigen (red) and GFP (green) along with DAPI (blue) at 12 hours PI with either HSV-1 KOS or HSV-1 ICP4- virus. A and B are representative figures of two experiments, n?=?3/group/experiment. Fold induction values were normalized to VEGF-A levels in uninfected, vehicle-treated controls. Panels C and D are representative of two experiments, n?=?3/group/experiment. Panel E is a summary of two experiments, n?=?6/group. Panel F is representative of two experiments with an n?=?3/group/experiment. Bars denote mean ± SEM. (** p<0.01, *<0.05, NSD?=?non-significant difference).

From: Wuest T, Zheng M, Efstathiou S, Halford WP, Carr DJJ (2011)
The Herpes Simplex Virus-1 Transactivator Infected Cell Protein-4 Drives VEGF-A Dependent Neovascularization.
PLoS Pathog 7(10): e1002278.

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  • Goat anti Green Fluorescent Protein
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  • Product Type
    Polyclonal Antibody
  • Isotype
    Polyclonal IgG
1 Formats Available
    Product CodeApplicationsDatasheetMSDSPack SizeList PriceQuantity
    AHP975E, IF, WBdatasheet pdfdatasheet pdf0.1 mg
    AHP975
    Summary
    Secondary Antibodies
    Negative Isotype Controls
    Useful Reagents
    Positive Controls
    Histology Controls
    More Images
    References
    Reviews
    -
    • Goat anti green fluorescent protein antibody recognizes green fluorescent protein (GFP), a 27kD protein which is derived from the jellyfish Aequorea victoria. GFP fluoresces green (509nm) when excited by blue light (395nm) and is commonly used as a marker of gene expression. The antibody recognises wild type GFP, but will also detect recombinant and enhanced forms of GFP.
    • Intended Use
    • Product Form
      Purified IgG - liquid
    • Reconstitution
    • Preparation
    • Antiserum Preparation
      Antisera to green fluorescent protein (GFP) were raised by repeated immunisations of goat with highly purified antigen. Purified IgG was prepared from whole serum by affinity chromatography.
    • Preservative Stabilisers
      0.01%Sodium Azide
    • Immunogen
      Fusion protein corresponding to full length GFP derived from Aequorea victoria
    • Purity
    • Approx. Protein Concentrations
      IgG concentration 1.0 mg/ml
    • Reagents In The Kit
    • Preparing The Antibody
    • Test Principle
    • Buffer Solution
      Phosphate buffered saline
    • Storage
      Store at +4oC or at -20oC if preferred.
      Storage in frost-free freezers is not recommended.
      This product should be stored undiluted. This product is photosensitive and should be protected from light.
      Avoid repeated freezing and thawing as this may denature the antibody. Should this product contain a precipitate we recommend microcentrifugation before use.
    • Shelf Life
      18 months from date of despatch.
    • GO Terms
      protein-chromophore linkage
      generation of precursor metabolites and energy
      bioluminescence
    • UniProt
    • Acknowledgements
    • Regulatory
      For research purposes only
    • This product has been reported to work in the following applications. This information is derived from testing within our laboratories, peer-reviewed publications or personal communications from the originators. Please refer to references indicated for further information. For general protocol recommendations, please visit the antibody protocols page.

    • Application NameYesNoMin DilutionMax Dilution
      ELISA
      Immunofluorescence
      Western Blotting

    • Where this antibody has not been tested for use in a particular technique this does not necessarily exclude its use in such procedures. Suggested working dilutions are given as a guide only. It is recommended that the user titrates the antibody for use in their own system using appropriate negative/positive controls.
    • Technical Advice
    • Recommended Protocol
    • ELISA
    • Immunohistology
    • Histology Positive Control Tissue
    • Immunofluorescence
    • Western Blotting
    • Instructions For Use

    Additional Green Fluorescent Protein Antibody Formats

    Formats Applications Sizes available
    Green Fluorescent Protein Antibody : Purified E, IF, WB 0.1 mg
    • Copyright © 2016 Bio-Rad

    Recommended Secondary Antibody

      DescriptionProduct CodePack SizeApplicationsList PriceQuantity
      Rabbit anti Goat IgG (Fc):FITCSTAR122F1 mgF
      STAR122F
      Rabbit anti Goat IgG (Fc):HRPSTAR122P1 mgC, E, WB
      STAR122P

      Recommended Negative Isotype Control

        Useful Reagents

          DescriptionProduct CodePack SizeApplicationsList PriceQuantity
          Rabbit anti Yellow Fluorescent ProteinAHP298350 µgWB
          AHP2983
          Rabbit anti Blue Fluorescent ProteinAHP298550 µgWB
          AHP2985
          Rabbit anti Cyan Fluorescent ProteinAHP298650 µgWB
          AHP2986
          Rabbit anti Red Fluorescent ProteinAHP298750 µgWB
          AHP2987

          Recommended Positive Controls

            Histology Controls

              • Application NameReference Images
                Western Blotting

              References

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